Re: IHC on bird tissue

From:AI Histology

At 09:39 AM 8/16/02 -0600, Jan Shivers wrote:
>Nonspecific staining problem in birds...
>I have recently begun to do IHC on bird tissue, and have come across a
>problem on certain cases where the inflammatory cells on the negative
>control stain as well.  (This is never a problem with any other mammalian
>species on which I've done IHC.)  Is there something unique or unusual about
>the avian hematopoietic system that requires an adjustment or change to the
>normal blocking protocols?  I've tried extending my 3% hydrogen peroxide
>quenching step to 30 minutes, but this doesn't eradicate the problem.  (I
>also use a 10% normal serum protein block prior to the primary antibody, and
>use the DAKO EnVision detection system with 2% normal avian serum in the
>EnVision reagent.)
>Helful hints would be MUCH APPRECIATED.
>Jan Shivers
>IHC Lab
>Univ. of Minnesota Vet. Diag. Lab

Hi Jan,

You might want to use methanol to make up your 3% hydrogen peroxide.
Although this is not
recommended for specimens where cell markers are to be stained. Also you
could try using
the peroxidase blocking reagent from Dako.  Good luck! 
Scott Fox H.T.(ASCP)QIHC
Ohio Department of Agriculture
Division of Animal Industry
Department Histology
8995 East Main St.

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